Numerical modelling of surface aeration and N2O emission in biological water resource recovery.

Biokinetic modelling of N2O production and emission has been extensively studied in the past fifteen years. In contrast, the physical-chemical hydrodynamics of activated sludge reactor design and operation, and their impact on N2O emission, is less well understood. This study addresses knowledge gaps related to the systematic identification and calibration of computational fluid dynamic (CFD) simulation models. Additionally, factors influencing reliable prediction of aeration and N2O emission in surface aerated oxidation ditch-type reactor types are evaluated. The calibrated model accurately predicts liquid sensor measurements obtained in the Lynetten Water Resource Recovery Facility (WRRF), Denmark. Results highlight the equal importance of design and operational boundary conditions, alongside biokinetic parameters, in predicting N2O emission. Insights into the limitations of calibrating gas mass-transfer processes in two-phase CFD models of surface aeration systems are evaluated.

Exploring the effects of intermittent aeration on the performance of nitrifying membrane-aerated biofilm reactors.

Membrane-aerated biofilm reactors (MABRs) are an emerging technology for nutrient removal; however, a trade-off remains between their removal rate and oxygen transfer efficiency. This study compares nitrifying flow-through MABRs operated under continuous and intermittent aeration modes at mainstream wastewater ammonia levels. The intermittently-aerated MABRs maintained maximal nitrification rates, including under conditions allowing the oxygen partial pressure on the gas side of the membrane to considerably drop during the no-aeration period. Nitrous oxide emissions of all reactors were comparable and amounted to approximately 20 % of the converted ammonia. Intermittent aeration increased the transformation rate constant of atenolol, yet did not affect the removal of sulfamethoxazole. Seven additional trace organic chemicals were not biodegraded by any of the reactors. The ammonia-oxidizing bacteria in the intermittently-aerated MABRs were dominated by Nitrosospira, previously shown to be abundant at low oxygen concentrations and provide reactor stability under changing conditions. Our findings indicate that intermittently-aerated flow-through MABRs can achieve high nitrification rates and oxygen transfer efficiencies, highlighting the possible implications of air supply discontinuity on nitrous oxide emissions and trace organic chemical biotransformation.

Critical evaluation of different mass transfer equations to model N2O emissions from water resource recovery facilities with diffuse aeration.

N2O measurements by liquid sensors in aerated tanks are an input to gas-liquid mass-transfer models for the prediction of N2O off-gas emissions. The prediction of N2O emissions from Water Resource Recovery Facilities (WRRFs) was evaluated by three different mass-transfer models using Benchmark Simulation Model 1 (BSM1) as a reference model. Inappropriate selection of mass-transfer model may result in miscalculation of carbon footprints based on soluble N2O online measurements. The film theory considers a constant mass-transfer expression, while more complex models suggest that emissions are affected by the aeration type, efficiency, and tank design characteristics. The differences among model predictions were 10-16% at dissolved oxygen (DO) concentration of 0.6 g/m3, when biological N2O production was the highest, while the flux of N2O was 20.0-24 kg N2O-N/d. At lower DO, the nitrification rate was low, while at DO higher than 2 g/m3, the N2O production was reduced leading to higher rates of complete nitrification and a flux of 5 kg N2O-N/d. The differences increased to 14-26% in deeper tanks, due to the pressure assumed in the tanks. The predicted emissions are also affected by the aeration efficiency when KLaN2O depends on the airflow instead of the KLaO2. Increasing the nitrogen loading rate under DO concentration of 0.50-0.65 g/m3 increased the differences in predictions by 10-20% in both alpha 0.6 and 1.2. A sensitivity analysis indicated that the selection of different mass-transfer models did not affect the selection of biochemical parameters for N2O model calibration.

Efficient management of the nitritation-anammox microbiome through intermittent aeration: absence of the NOB guild and expansion and diversity of the NOx reducing guild suggests a highly reticulated nitrogen cycle.

Obtaining efficient autotrophic ammonia removal (aka partial nitritation-anammox, or PNA) requires a balanced microbiome with abundant aerobic and anaerobic ammonia oxidizing bacteria and scarce nitrite oxidizing bacteria. Here, we analyzed the microbiome of an efficient PNA process that was obtained by sequential feeding and periodic aeration. The genomes of the dominant community members were inferred from metagenomes obtained over a 6 month period. Three Brocadia spp. genomes and three Nitrosomonas spp. genomes dominated the autotrophic community; no NOB genomes were retrieved. Two of the Brocadia spp. genomes lacked the genomic potential for nitrite reduction. A diverse set of heterotrophic genomes was retrieved, each with genomic potential for only a fraction of the denitrification pathway. A mutual dependency in amino acid and vitamin synthesis was noted between autotrophic and heterotrophic community members. Our analysis suggests a highly-reticulated nitrogen cycle in the examined PNA microbiome with nitric oxide exchange between the heterotrophs and the anammox guild.

Mainstream short-cut N removal modelling: current status and perspectives.

This work gives an overview of the state-of-the-art in modelling of short-cut processes for nitrogen removal in mainstream wastewater treatment and presents future perspectives for directing research efforts in line with the needs of practice. The modelling status for deammonification (i.e., anammox-based) and nitrite-shunt processes is presented with its challenges and limitations. The importance of mathematical models for considering N2O emissions in the design and operation of short-cut nitrogen removal processes is considered as well. Modelling goals and potential benefits are presented and the needs for new and more advanced approaches are identified. Overall, this contribution presents how existing and future mathematical models can accelerate successful full-scale mainstream short-cut nitrogen removal applications.

Chronic effects of cerium dioxide nanoparticles on biological nitrogen removal and nitrous oxide emission: Insight into impact mechanism and performance recovery potential.

The influence of cerium dioxide nanoparticles (CeO2 NPs) on biological nitrogen removal and associated nitrous oxide (N2O) emission has seldom been addressed yet. Herein, the chronic effect of CeO2 NPs on the nitrogen transformation processes during wastewater treatment and the impacted system's self-recovery potential after CeO2 NP stress removal were investigated. CeO2 NP of 10-50 mg/L induced significant declines of the ammonia nitrogen (NH4+-N) and the total nitrogen removal efficiencies, but triggered the nitrite accumulation and the N2O emission. The N2O reductase (NOS) activity was negatively correlated with the N2O emission level, and the inhibition of NOS activity under CeO2 NP stress was probably due to the depressions of the sludge denitrifiers' metabolic activities. The NH4+-N removal efficiency was successfully regained after the recovery period although the N2O emission level was still higher than the pre-exposure period, which was probably due to the residual CeO2 NPs inside the activated sludge.

Insights into chronic zinc oxide nanoparticle stress responses of biological nitrogen removal system with nitrous oxide emission and its recovery potential.

The nitrogen transformation performances and greenhouse gas nitrous oxide (N2O) emissions in a sequencing batch reactor under chronic exposure to zinc oxide nanoparticles (ZnO NPs) were quantified and the system's self-recovery potentials were assessed. ZnO NPs posed a dose-dependent depression effect on the removal efficiencies of ammonia nitrogen (NH4+-N) and total nitrogen (TN), and the N2O emissions. The suppressed N2O emissions had a positive relationship with the activity ratios of nitrite/NO reductases and N2O reductase, and were expected to be caused by the inhibited heterotrophic denitrification process. The inhibition of glucose metabolism key enzymes and electron transport chain activities would be responsible for the heterotrophic denitrification performances deterioration. Furthermore, the removal efficiencies of NH4+-N and TN were recovered to control levels through the nitrite-shunt. However, the N2O emission increased significantly above the control during the recovery period mainly due to the irreversibility of the depressed nitrite oxidation activities.

Modeling Denitrification as an Electric Circuit Accurately Captures Electron Competition between Individual Reductive Steps: The Activated Sludge Model-Electron Competition Model.

Heterotrophic denitrification consists of the four-step sequential reduction of nitrate to dinitrogen gas over nitrite, nitric oxide, and nitrous oxide. Oxidation processes, commonly of organic compounds, provide the electrons needed for the sequential reaction steps. The intracellular electron distribution is a competitive process among the four reduction steps. In this study, a model describing organic carbon oxidation and four-step denitrification through electron competition is proposed [Activated Sludge Model-Electron Competition (ASM-EC)]. The model describes denitrification rates as an analogy to how current intensity varies through a parallel set of resistors in electric circuits. The ASM-EC model was calibrated with data from batch experiments with heterotrophic denitrifying communities, where reduction of mixtures of nitrogen oxides was monitored, while different carbon sources were supplied in excess. The carbon sources included methanol, ethanol, acetate, and their ternary mixture. The electron distribution preference and electron uptake rates varied between the carbon sources and were captured by the model structure for most of the experiments. The ASM-EC model uses fewer parameters compared to existing state-of-the-art denitrification models and performed equally well in the tested scenarios. We advocate the use of this model for denitrification in the activated sludge model, which can easily be integrated in existing model structures, because it provides a parsimonious description of electron competition during denitrification.

Regulation of key N2O production mechanisms during biological water treatment.

Nitrous oxide (N2O) is a potent greenhouse gas emitted during biological treatment of residual waters and can contribute significantly to the carbon footprint of the overall treatment, potentially offsetting energy-positive strategies. N2O production is mediated by three known biological pathways and through abiotic reactions, driven by biologically generated substances such as hydroxylamine and nitrite. The contributions of these different mechanism are determined by the environmental conditions and the resident microbial community. The newly discovered phenotypic diversity among aerobic ammonia oxidizers and the modularity of denitrifying pathway determines N2O emissions. Isotopic methods can be used to quantify N2O production pathways in water treatment systems, and mechanistic models can already predict N2O emissions, but limitations on their accuracy and precision still exist.

The effect of pH on N2O production in intermittently-fed nitritation reactors.

The effect of pH on nitrous oxide (N2O) production rates was quantified in an intermittently-fed lab-scale sequencing batch reactor performing high-rate nitritation. N2O and other nitrogen (N) species (e.g. ammonium (NH4+), nitrite, hydroxylamine and nitric oxide) were monitored to identify in-cycle dynamics and determine N conversion rates at controlled pH set-points (6.5, 7, 7.5, 8 and 8.5). Operational conditions and microbial compositions remained similar during long-term reactor-scale pH campaigns. The specific ammonium removal rates and nitrite accumulation rates varied little with varying pH levels (p > 0.05). The specific net N2O production rates and net N2O yield of NH4+ removed (ΔN2O/ΔNH4+) increased up to seven-fold from pH 6.5 to 8, and decreased slightly with further pH increase to 8.5 (p < 0.05). Best-fit model simulations predicted nitrifier denitrification as the dominant N2O production pathway (≥87% of total net N2O production) at all examined pH. Our study highlights the effect of pH on biologically mediated N2O emissions in nitrogen removal systems and its importance in the design of N2O mitigation strategies.

Abiotic Nitrous Oxide (N2O) Production Is Strongly pH Dependent, but Contributes Little to Overall N2O Emissions in Biological Nitrogen Removal Systems.

Hydroxylamine (NH2OH) and nitrite (NO2-), intermediates during the nitritation process, can engage in chemical (abiotic) reactions that lead to nitrous oxide (N2O) generation. Here, we quantify the kinetics and stoichiometry of the relevant abiotic reactions in a series of batch tests under different and relevant conditions, including pH, absence/presence of oxygen, and reactant concentrations. The highest N2O production rates were measured from NH2OH reaction with HNO2, followed by HNO2 reduction by Fe2+, NH2OH oxidation by Fe3+, and finally NH2OH disproportionation plus oxidation by O2. Compared to other examined factors, pH had the strongest effect on N2O formation rates. Acidic pH enhanced N2O production from the reaction of NH2OH with HNO2 indicating that HNO2 instead of NO2- was the reactant. In departure from previous studies, we estimate that abiotic N2O production contributes little (< 3% of total N2O production) to total N2O emissions in typical nitritation reactor systems between pH 6.5 and 8. Abiotic contributions would only become important at acidic pH (≤ 5). In consideration of pH effects on both abiotic and biotic N2O production pathways, circumneutral pH set-points are suggested to minimize overall N2O emissions from nitritation systems.

The pH dependency of N-converting enzymatic processes, pathways and microbes: effect on net N2 O production.

Nitrous oxide (N2 O) is emitted during microbiological nitrogen (N) conversion processes, when N2 O production exceeds N2 O consumption. The magnitude of N2 O production vs. consumption varies with pH and controlling net N2 O production might be feasible by choice of system pH. This article reviews how pH affects enzymes, pathways and microorganisms that are involved in N-conversions in water engineering applications. At a molecular level, pH affects activity of cofactors and structural elements of relevant enzymes by protonation or deprotonation of amino acid residues or solvent ligands, thus causing steric changes in catalytic sites or proton/electron transfer routes that alter the enzymes' overall activity. Augmenting molecular information with, e.g., nitritation or denitrification rates yields explanations of changes in net N2 O production with pH. Ammonia oxidizing bacteria are of highest relevance for N2 O production, while heterotrophic denitrifiers are relevant for N2 O consumption at pH > 7.5. Net N2 O production in N-cycling water engineering systems is predicted to display a 'bell-shaped' curve in the range of pH 6.0-9.0 with a maximum at pH 7.0-7.5. Net N2 O production at acidic pH is dominated by N2 O production, whereas N2 O consumption can outweigh production at alkaline pH. Thus, pH 8.0 may be a favourable pH set-point for water treatment applications regarding net N2 O production.

Calibration of the comprehensive NDHA-N2O dynamics model for nitrifier-enriched biomass using targeted respirometric assays.

The NDHA model comprehensively describes nitrous oxide (N2O) producing pathways by both autotrophic ammonium oxidizing and heterotrophic bacteria. The model was calibrated via a set of targeted extant respirometric assays using enriched nitrifying biomass from a lab-scale reactor. Biomass response to ammonium, hydroxylamine, nitrite and N2O additions under aerobic and anaerobic conditions were tracked with continuous measurement of dissolved oxygen (DO) and N2O. The sequential addition of substrate pulses allowed the isolation of oxygen-consuming processes. The parameters to be estimated were determined by the information content of the datasets using identifiability analysis. Dynamic DO profiles were used to calibrate five parameters corresponding to endogenous, nitrite oxidation and ammonium oxidation processes. The subsequent N2O calibration was not significantly affected by the uncertainty propagated from the DO calibration because of the high accuracy of the estimates. Five parameters describing the individual contribution of three biological N2O pathways were estimated accurately (variance/mean < 10% for all estimated parameters). The NDHA model response was evaluated with statistical metrics (F-test, autocorrelation function). The 95% confidence intervals of DO and N2O predictions based on the uncertainty obtained during calibration are studied for the first time. The measured data fall within the 95% confidence interval of the predictions, indicating a good model description. Overall, accurate parameter estimation and identifiability analysis of ammonium removal significantly decreases the uncertainty propagated to N2O production, which is expected to benefit N2O model discrimination studies and reliable full scale applications.

Low nitrous oxide production through nitrifier-denitrification in intermittent-feed high-rate nitritation reactors.

Nitrous oxide (N2O) production from autotrophic nitrogen conversion processes, especially nitritation systems, can be significant, requires understanding and calls for mitigation. In this study, the rates and pathways of N2O production were quantified in two lab-scale sequencing batch reactors operated with intermittent feeding and demonstrating long-term and high-rate nitritation. The resulting reactor biomass was highly enriched in ammonia-oxidizing bacteria, and converted ∼93 ± 14% of the oxidized ammonium to nitrite. The low DO set-point combined with intermittent feeding was sufficient to maintain high nitritation efficiency and high nitritation rates at 20-26 °C over a period of ∼300 days. Even at the high nitritation efficiencies, net N2O production was low (∼2% of the oxidized ammonium). Net N2O production rates transiently increased with a rise in pH after each feeding, suggesting a potential effect of pH on N2O production. In situ application of 15N labeled substrates revealed nitrifier denitrification as the dominant pathway of N2O production. Our study highlights operational conditions that minimize N2O emission from two-stage autotrophic nitrogen removal systems.

Intermittent Aeration Suppresses Nitrite-Oxidizing Bacteria in Membrane-Aerated Biofilms: A Model-Based Explanation.

Autotrophic ammonium oxidation in membrane-aerated biofilm reactors (MABRs) can make treatment of ammonium-rich wastewaters more energy-efficient, especially within the context of short-cut ammonium removal. The challenge is to exclusively enrich ammonium-oxidizing bacteria (AOB). To achieve nitritation, strategies to suppress nitrite-oxidizing bacteria (NOB) are needed, which are ideally grounded on an understanding of underlying mechanisms. In this study, a nitrifying MABR was operated under intermittent aeration. During eight months of operation, AOB dominated, while NOB were suppressed. On the basis of dissolved oxygen (DO), ammonium, nitrite, and nitrate profiles within the biofilm and in the bulk, a 1-dimensional nitrifying biofilm model was developed and calibrated. The model was utilized to explore the potential mechanisms of NOB suppression associated with intermittent aeration, considering DO limitation, direct pH effects on enzymatic activities, and indirect pH effects on activity via substrate speciation. The model predicted strong periodic shifts in the spatial gradients of DO, pH, free ammonia, and free nitrous acid, associated with aerated and nonaerated phases. NOB suppression during intermittent aeration was mostly explained by periodic inhibition caused by free ammonia due to periodic transient pH upshifts. Dissolved oxygen limitation did not govern NOB suppression. Different intermittent aeration strategies were then evaluated for nitritation success in intermittently aerated MABRs: both aeration intermittency and duration were effective control parameters.

Heterotrophs are key contributors to nitrous oxide production in activated sludge under low C-to-N ratios during nitrification-Batch experiments and modeling.

Nitrous oxide (N2 O), a by-product of biological nitrogen removal during wastewater treatment, is produced by ammonia-oxidizing bacteria (AOB) and heterotrophic denitrifying bacteria (HB). Mathematical models are used to predict N2 O emissions, often including AOB as the main N2 O producer. Several model structures have been proposed without consensus calibration procedures. Here, we present a new experimental design that was used to calibrate AOB-driven N2 O dynamics of a mixed culture. Even though AOB activity was favoured with respect to HB, oxygen uptake rates indicated HB activity. Hence, rigorous experimental design for calibration of autotrophic N2 O production from mixed cultures is essential. The proposed N2 O production pathways were examined using five alternative process models confronted with experimental data inferred. Individually, the autotrophic and heterotrophic denitrification pathway could describe the observed data. In the best-fit model, which combined two denitrification pathways, the heterotrophic was stronger than the autotrophic contribution to N2 O production. Importantly, the individual contribution of autotrophic and heterotrophic to the total N2 O pool could not be unambiguously elucidated solely based on bulk N2 O measurements. Data on NO would increase the practical identifiability of N2 O production pathways. Biotechnol. Bioeng. 2017;114: 132-140. © 2016 Wiley Periodicals, Inc.

Aeration strategies to mitigate nitrous oxide emissions from single-stage nitritation/anammox reactors.

Autotrophic nitrogen removal is regarded as a resource efficient process to manage nitrogen-rich residual streams. However, nitrous oxide emissions of these processes are poorly documented and strategies to mitigate emissions unknown. In this study, two sequencing batch reactors performing single-stage nitritation/anammox were operated under different aeration strategies, gradually adjusted over six months. At constant but limiting oxygen loading, synthetic reject water was fed (0.75 g-N/L · d) and high nitrogen removal efficiencies (83 ± 5 and 88 ± 2%) obtained. Dynamics of liquid phase nitrous (N2O) and nitric oxide (NO) concentrations were monitored and N2O emissions calculated. Significant decreases in N2O emissions were obtained when the frequency of aeration was increased while maintaining a constant air flow rate (from >6 to 1.7% ΔN2O/ΔTN). However, no significant effect on the emissions was noted when the duration of aeration was increased while decreasing air flow rate (10.9 ± 3.2% ΔN2O/ΔTN). The extant ammonium oxidation activity (mgNH4(+)-N/gVSS · min) positively correlated with the specific N2O production rate (mgN2O-N/gVSS · min) of the systems. Operating under conditions where anaerobic exceeds aerobic ammonium oxidation activity is proposed to minimize N2O emissions from single-stage nitritation/anammox reactors; increasing the frequency of aeration cycling is an efficient way of obtaining those conditions.

Critical assessment of extracellular polymeric substances extraction methods from mixed culture biomass.

Extracellular polymeric substances (EPS) have a presumed determinant role in the structure, architecture, strength, filterability, and settling behaviour of microbial solids in biological wastewater treatment processes. Consequently, numerous EPS extraction protocols have recently been published that aim to optimize the trade off between high EPS recovery and low cell lysis. Despite extensive efforts, the obtained results are often contradictory, even when analysing similar biomass samples and using similar experimental conditions, which greatly complicates the selection of an extraction protocol. This study presents a rigorous and critical assessment of existing physical and chemical EPS extraction methods applied to mixed-culture biomass samples (nitrifying, nitritation-anammox, and activated sludge biomass). A novel fluorescence-based method was developed and calibrated to quantify the lysis potential of different EPS extraction protocols. We concluded that commonly used methods to assess cell lysis (DNA concentrations or G6PDH activities in EPS extracts) do not correlate with cell viability. Furthermore, we discovered that the presence of certain chemicals in EPS extracts results in severe underestimation of protein and carbohydrate concentrations by using standard analytical methods. Keeping both maximum EPS extraction yields and minimal biomass lysis as criteria, it was identified a sonication-based extraction method as the best to determine and compare tightly-bound EPS fractions in different biomass samples. Protein was consistently the main EPS component in all analysed samples. However, EPS from nitrifying enrichments was richer in DNA, the activated sludge EPS had a higher content in humic acids and carbohydrates, and the nitritation-anammox EPS, while similar in composition to the nitrifier EPS, had a lower fraction of hydrophobic biopolymers. In general, the easily-extractable EPS fraction was more abundant in carbohydrates and humic substances, while DNA could only be found in tightly bound EPS fractions. In conclusion, the methodology presented herein supports the rational selection of analytical tools and EPS extraction protocols in further EPS characterization studies.